Self-Assembly and Protein–Protein Interactions between the SV40 Capsid Proteins Produced in Insect Cells

Sandalon, Ziv; Oppenheim, Ariella

doi:10.1006/viro.1997.8796

Cited by 74 publications

(64 citation statements)

References 31 publications

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“…Dithiothreitol, PMSF and the protease inhibitor cocktail were added to the buffer immediately before use. 14,15 Nuclear extract concentration was measured using the BCA Protein Assay protocol (Pierce, Rockford, IL). The nuclear extract was kept for more than a year at À201C.…”

Section: Preparation Of Nuclear Extracts From Sf9 Cellsmentioning

confidence: 99%

SV40 Pseudovirion gene delivery of a toxin to treat human adenocarcinomas in mice

et al. 2006

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SV40 vectors packaged in vitro (pseudovirions) are an efficient delivery system for plasmids up to 17.7 kb, with or without SV40 sequences. A truncated Pseudomonas exotoxin gene (PE38) was delivered into various human cells (HeLa, KB-3-1, human lymphoblastoids, and erythroleukemia cells), in vitro using pseudovirions. The number of viable cells was reduced significantly in the PE38-transduced cells. Human KB adenocarcinomas growing in mice were treated with intratumoral injection of PE38 packaged in vitro, and tumor size decreased significantly. Intraperitoneal treatments were as effective in reducing tumor size as intratumoral treatments. To check the viability of mock-or PE38-treated mice, every 4 days they were weighed, their blood was tested, and various tissues were screened for pathology. All parameters showed that the in vitro-packaged vectors, injected into tumors or intraperitoneally, caused no abnormalities in mice. The combined treatment of doxorubicin with in vitro-packaged PE38 reduced tumor size slightly more than each of the treatments separately. However, the combined treatment did not cause the weight loss seen with doxorubicin alone. These results indicate that SV40 in vitro packaging is an effective system for cancer gene delivery using two different routes of injection and in combination with chemotherapy.

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Section: Preparation Of Nuclear Extracts From Sf9 Cellsmentioning

confidence: 99%

SV40 Pseudovirion gene delivery of a toxin to treat human adenocarcinomas in mice

et al. 2006

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“…5B). As a source of VP1 we used nuclear extracts of Sf9 cells infected with a recombinant baculovirus that expresses VP1 (30). The reaction was performed in the presence of double-stranded poly(dI-dC) to compete for the nonspecific binding activity of VP1 (33) and at a low concentration of Sp1, conditions at which Sp1 alone and VP1 alone each bound poorly to the DNA probe (lanes 2 and 3 and lane 7, respectively).…”

Section: Vol 76 2002mentioning

confidence: 99%

“…SV40 virions and pseudovirions can be prepared in vitro. The three capsid proteins VP1, VP2, and VP3, produced as recombinant proteins in insect cells, localize to the nucleus where they assemble spontaneously to form virus-like particles (30). Virus-like particles are capable of incorporating in vitro either SV40 or plasmid DNA, producing infectious particles (29).…”

Section: Vol 76 2002mentioning

confidence: 99%

Cellular Transcription Factor Sp1 Recruits Simian Virus 40 Capsid Proteins to the Viral Packaging Signal, ses

Gordon‐Shaag

Ben-nun-Shaul

Roitman

et al. 2002

J Virol

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Simian virus 40 (SV40) capsid assembly occurs in the nucleus. All three capsid proteins bind DNA nonspecifically, raising the dilemma of how they attain specificity to the SV40 minichromosome in the presence of a large excess of genomic DNA. The SV40 packaging signal, ses, which is required for assembly, is composed of multiple DNA elements that bind transcription factor Sp1. Our previous studies showed that Sp1 participates in SV40 assembly and that it cooperates in DNA binding with VP2/3. We hypothesized that Sp1 recruits the capsid proteins to the viral minichromosome, conferring upon them specific DNA recognition. Here, we have tested the hypothesis. Computer analysis showed that the combination of six tandem GC boxes at ses is not found at cellular promoters and therefore is unique to SV40. Cooperativity in DNA binding between Sp1 and VP2/3 was not abolished at even a 1,000-fold excess of cellular DNA, providing strong support for the recruitment hypothesis. Sp1 also binds VP1 and cooperates with VP1 in DNA binding. VP1 pentamers (VP1 5 ) avidly interact with VP2/3, utilizing the same VP2/3 domain as described for polyomavirus. We conclude that VP1 5 -VP2/3 building blocks are recruited by Sp1 to ses, where they form the nucleation center for capsid assembly. By this mechanism the virus ensures that capsid formation is initiated at a single site around its minichromosome.

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“…Such structures were originally described for mouse polyoma virus coat protein, VP1, [3][4][5] and it has been shown that a number of different viral coat proteins can behave in a similar manner. [6][7][8][9][10][11][12][13][14][15][16][17] VLPs retain their ability to package DNA. Particles composed of the coat protein packaged with heterologous DNA have been isolated from cells in which the recombinant protein is syn- somes or calcium phosphate precipitates.…”

Section: Introductionmentioning

confidence: 99%

Virus-like gene transfer into cells mediated by polyoma virus pseudocapsids

et al. 2000

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Mouse polyoma virus-like particles (or pseudocapsids) are composed solely of recombinant viral coat protein. They can interact with DNA and transport it to cells, resulting in gene expression both in tissue culture and in mice. We demonstrate that DNA transfer in vitro depends on partial packaging of DNA within the virus-like capsid. Cell surface sialic acid residues and an intact microtubule network, required for viral infectivity, are also necessary for pseudocapsidmediated gene expression from heterologous DNA. Thus, gene delivery in this system requires pathways utilised by polyoma virions, rather than proceeding via the 'nonspecific' endosomal route typical of nonviral systems such as lipo-

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Self-Assembly and Protein–Protein Interactions between the SV40 Capsid Proteins Produced in Insect Cells

Cited by 74 publications

References 31 publications

SV40 Pseudovirion gene delivery of a toxin to treat human adenocarcinomas in mice

SV40 Pseudovirion gene delivery of a toxin to treat human adenocarcinomas in mice

Cellular Transcription Factor Sp1 Recruits Simian Virus 40 Capsid Proteins to the Viral Packaging Signal, ses

Virus-like gene transfer into cells mediated by polyoma virus pseudocapsids

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